Human and Animal Fascioliasis in Iran: Systematic Review and Meta-analysis.

OBJECTIVE: The aim of this study was to investigate the prevalence of fascioliasis in humans, cattle, sheep, and goats, done by a systematic review in Iran. METHODS: Fifty articles were extracted including Iranian papers such as Google scholar, Magiran, Iran Medex, SID, and Pubmed. Out of these, 21 articles were selected for meta-analysis. Essential information for meta-analysis was extracted from papers and archived in Excel software for calculating by statistical analysis. The variance of each study was obtained using the binomial distribution. Heterogeneity of the studies was surveyed using I2 index. Data were analyzed using a random effect model. RESULTS: Of 21 collected papers, 1,275,506 samples from cow (507,152), sheep (454,882), goat (207,925), and human (105,547) had been surveyed in Iran. Eight studies were conducted on humans and 13 on animals. The prevalence rate obtained in humans was 3% with a confidence interval (CI) of 95% (1%-7%). Prevalence rate obtained in cows, sheep, and goats was 13% with CI of 95% (10%-16%). The highest level of prevalence was reported from cities in the North in animals with a prevalence of 14%. The highest level of prevalence was reported from Gilan in humans with a prevalence of 0.1%. CONCLUSION: The prevalence of the Fascioliasis in Iran has reduced in recent years, But the importance of the disease has not reduced and there is a possibility of an epidemic. Furthermore, in many cities of Iran, there is no study on the disease.

What are Hidden Facts behind Intestinal Parasitic Infections in Ilam City?

BACKGROUND: Intestinal parasitic infections are the one of the most common health problems in developing countries. OBJECTIVE: A number of patients die annually due to complications caused by these parasites.Therefore, the aim of this study was to investigate the rate and type of parasitic infections, determine the factors affecting them in Ilam city and also provide strategies to prevent them.In this descriptive cross-sectional study conducted in one of the Ilam labs in 2016, 417 stool specimens were randomly collected. All specimens were examined using direct and ethanol formaldehyde.Suspect specimens were examined using Trichrom staining. Demographic information was also recorded in a questionnaire, and finally the results were analyzed using statistical software SPSS 20.The data were then compared with Chi-square test. RESULTS: Out of the 417 patients examined, 59 (14.1%) were infected with intestinal parasites. The type of parasitic infection in 9.4% was Blastocystis hominis, 3.6% Entamoeba coli, 0.5% Entamoeba histolytica, 0.5% Giardia and 0.2% Trichomonas hominis. CONCLUSION: Despite the improvement of public health, parasitic infections are still considered as one of the health problems in the city of Ilam. Therefore, proper planning, public health education, raising the level of health in the area and the provision of safe drinking water are some of the ways to reduce parasitic infections in the region.

Omega-3 fatty acids, EPA and DHA induce apoptosis and enhance drug sensitivity in multiple myeloma cells but not in normal peripheral mononuclear cells.

The n-3 polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been shown to enhance the effect of chemotherapeutic drugs in clinical studies in cancer patients and to induce apoptotic tumor cell death in vitro. Until now, EPA and DHA have never been investigated in multiple myeloma (MM). Human myeloma cells (L363, OPM-1, OPM-2 and U266) and normal peripheral blood mononuclear cells were exposed to EPA and DHA, and effects on mitochondrial function and apoptosis, caspase-3 activation, gene expression and drug toxicity were measured. Exposure to EPA and DHA induced apoptosis and increased sensitivity to bortezomib in MM cells. Importantly, they did not affect viability of normal human peripheral mononuclear cells. Messenger RNA expression arrays showed that EPA and DHA modulated genes involved in multiple signaling pathways including nuclear factor (NF) κB, Notch, Hedgehog, oxidative stress and Wnt. EPA and DHA inhibited NFκB activity and induced apoptosis through mitochondrial perturbation and caspase-3 activation. Our study suggests that EPA and DHA induce selective cytotoxic effects in MM and increase sensitivity to bortezomib and calls for further exploration into a potential application of these n-3 polyunsaturated fatty acids in the therapy of MM.

Toll-like receptors in human multiple myeloma: new insight into inflammation-related pathogenesis.

Multiple myeloma (MM) is a clonal neoplasm characterized by expansion of malignant plasma cells in the bone marrow causing various complications including osteolytic lesions and impaired immune function. It has recently been reported that human myeloma cells express multiple Toll-like receptors (TLRs), and their activation-induced functional responses show heterogeneity among cell lines and patient samples. TLRs are critical germ-line encoded molecules expressed in immune cells as well as in a variety of cancer cells. In multiple myeloma, they may induce cell growth and proliferation or promote cell death. In fact, our current knowledge of Toll-like receptor function has gone beyond their main function as triggers of innate and adaptive immune responses. Considering the essential role of bone marrow microenvironment components in myeloma tumor expansion, survival, invasion and drug resistance, TLR triggering may contribute to adhesion-induced or de novo drug resistance of MM cells. Future preclinical and clinical studies are needed to address if TLRs can be exploited as novel therapeutic targets for MM.

Stimulation of Toll-like receptor-1/2 combined with Velcade increases cytotoxicity to human multiple myeloma cells.

An increasing body of evidence supports the important role of adhesion to bone marrow microenvironment components for survival and drug resistance of multiple myeloma (MM) cells. Previous studies suggested that stimulation of Toll-like receptors by endogenous ligands released during inflammation and tissue damage may be pro-tumorigenic, but no studies have been performed in relation to modulation of cell adhesion and drug cytotoxicity. Here, we investigated the effect of TLR1/2 activation on adhesion of human myeloma cells to fibronectin, and their sensitivity to the proteasome inhibitor Velcade. It was found that TLR1/2 activation with Pam3CSK4 increased the cytotoxicity of Velcade in L363, OPM-2 and U266 human myeloma cells. This effect was not related to a decreased adhesion of the cells to fibronectin, but TLR1/2 activation stimulated the caspase-3 activity in Velcade-treated myeloma cells, which may be responsible for the enhanced cell death. Inhibitors of NF-κB and MAPK reduced the stimulatory effect. These findings indicate that TLR activation of MM cells could bypass protective effects of cell adhesion and suggest that TLR signaling may also have antitumorigenic potential.

Genotyping of Hydatid Cyst Isolated from Human and Domestic Animals in Ilam Province, Western Iran Using PCR-RFLP.

BACKGROUND: Hydatidosis or cystic hydatid disease is one of the most important diseases in human and animals. Identification of strains is important for improvement of control and prevention of disease. The aim of this study was to determine the strains isolated from human and domestic animals in Ilam Province, Iran, using PCR-RFLP method. METHODS: Respectively, 30 and 4 animal and human hydatid cysts were collected from different slaughterhouses and hospitals of the province. Protoscolices were separated and their DNA genome was extracted by extraction kit. rDNA-ITS1 of each isolated samples was duplicated by BD1(Forward) and 4s (Reverse) Primers. PCR products were studied by electrophoresis and then were digested using TaqI, HpaII, RsaI and AluI restriction enzymes. RFLP products were studied using electrophoresis on 1% agar gel. RESULT: A fragment of 1000bp was produced from amplification of rDNA-ITS1 of protoscolices using PCR method. After digestion of PCR product by AluI enzyme, 200bp and 800bp, by RsaI, 655bp and 345bp and by HpaII 700bp and 300bp sizes were obtained. TaqI enzyme had no change in fragment size and it remained 1000bp. Considering the method, Ilam strains was specified as E. granulosus sensu stricto (G1-G3). CONCLUSIONS: Although sheep strain (G1) is dominated in human and different animal in Iran and the world, but more efforts should be done to clarify the true genotype of Ilam strains specified as E. granulosus sensu stricto (G1-G3).

Seroepidemiological study of human hydatidosis in meshkinshahr district, ardabil province, iran.

BACKGROUND: The aim of this study was to conduct a sero-epidemiological survey in Meshkinshahr, Ardabil Province, northwestern Iran to detect the rate of hydatidosis in the city and nearby villages. Literature shows that no such study has been conducted so far. METHODS: Overall, 670 serum samples were collected from 194 males and 476 females from patients referred to different health centers of the region. All patients filled out a questionnaire and an informed consent. Sera were analyzed using indirect-ELISA test. Ten µg /ml antigens (Antigen B derived from hydatid cyst fluid), serum dilutions of 1:500 and conjugate anti-human coombs with 1:10000 dilutions were utilized to perform the test. Data analysis was conducted using SPSS software ver. 11.5. RESULTS: The seroprevalence of human hydatidosis was 1.79% by ELISA test in the region. This rate for females was 1.68% and males 2.6%, respectively. There was no significant difference as regards all factors studied and the seropositivity. According to job, farmers and ranchmen had the highest rate of infection as 3.17%. The sero-prevalence of infection was 2.6%% in illiterate people which showed the highest rate. As regards residency, urban life showed no significant difference with rural life (1.1% vs. 2.58%). Age group of 69-90 yr old, with 4.62% as prevalence had the highest rate of positivity. CONCLUSION: Obtained sero-prevalence of hydatidosis shows more or less a resemblance to other cities of Iran, although due to the specific condition of the city we expected more rate of sero-positivity.

Gene cloning, expression and serological evaluation of the 12-kDa antigen-B subunit from Echinococcus granulosus.

A 12-kDa subunit of antigen B from Echinococcus granulosus has recently been cloned, expressed and used in diagnostic ELISA to test human sera for evidence of cystic echinococcosis. The performance of the ELISA based on the recombinant antigen (rAgB) was compared with that of similar assays based on native antigen B (nAgB) or hydatid-cyst fluid. For the preparation of the rAgB, total RNA was extracted from Ec. granulosus protoscoleces so that antigen-B complementary DNA could be synthesised, amplified by PCR, and then cloned into the pQE30 expression vector. The recombinant plasmid was transformed in Escherichia coli and induced using isopropyl-beta-D-thiogalactopyrano-side. Bacterial samples were collected, lysed and then analysed by SDS-PAGE and western blotting. The recombinant protein was purified by affinity chromatography. Although the performance of the ELISA based on cyst fluid appeared identical to that of the assay based on the recombinant antigen (with a sensitivity, specificity, positive predictive value and negative predictive value of 96.0%, 97.0%, 97.2% and 95.5%, respectively), the corresponding results for the ELISA based on nAgB (98.6%, 100%, 100% and 98.5%) were slightly better. Despite this difference (which was not statistically significant), the comparative ease with which large quantities of the recombinant antigen could be produced make the antigen a potentially useful tool in the diagnosis of cystic echinococcosis.

Serological Evaluation of EgAgB16 kDa, a Recombinant Antigen from Echinococcus granulosus for Diagnosis of Human Hydatidosis.

BACKGROUND: Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test. METHODS: DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individuals (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5) and tuberculosis (n=5) were examined using this recombinant antigen. RESULTS: Recombinant protein was purified by affinity chromatography using His-Tag column. After purification, recombinant protein was confirmed by western blot analysis using His Tag monoclonal antibody or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for rAgB16. CONCLUSION: While the produced recombinant AgB16 kDa showed promising results in diagnosing human hydatidosis, but more investigations should be implemented to reach an accurate gold standard.